Peptides for diagnosing lyme disease
The present invention relates, e.g., to a composition comprising peptides represented by SEQ ID NO:1, or active variants thereof, wherein the peptides or active variants can bind specifically to an antibody induced by a causative agent of Lyme disease (a pathogenic Borrelia ), e.g. in a sample from a subject having Lyme disease. Compositions of the invention may comprise multiple peptides, from multiple proteins. Diagnostic kits comprising the peptides are described, as are diagnostic assays using the peptides.
1. A composition comprising a linked peptide, comprising
(a) a first peptide KDLKNKILKIKKEATGKGVLFEAFTGLKTG (SEQ ID NO: 1), or an active variant thereof wherein one or more of the amino acids of SEQ ID NO: 1 is substituted with an amino acid replacement, wherein the peptide or active variant specifically binds to an antibody against a pathogenic Borrelia;
covalently linked by a peptide linker to
(b) at least one additional peptide that specifically recognizes an antibody against the same or a different protein of the same or a different pathogenic Borrelia ; and
wherein the linked peptide is selected from the group consisting of:
(SEQ ID NO: 6)
KDLKNKILKIKKEATGKGVLFEAFTG
LKTG GGGMKKNDQIVAAIALRGVA,
(SEQ ID NO: 7)
KDLKNKILKIKKEATGKGVLFEAFTG
LKTG GGGPFILEAKVRATTVAE,
(SEQ ID NO: 8)
KDLKNKILKIKKEATGKGVLFEAFTG
LKTGGG NKTFNNLLKLTILVNGGGTI
LVNLLISCGLTGA,
(SEQ ID NO: 9)
KDLKNKILKIKKEATGKGVLFEAFTG
LKTG GGGNKTFNNLLKLTILVN,
(SEQ ID NO: 10)
DNVKNKILQIKEEAAKKGVNFKAFTG
TATG GGGMKKNDQIVAAIALRGVA,
(SEQ ID NO: 11)
DNVKNKILQIKEEAAKKGVNFKAFTG
TATG GGGPFILEAKVRATTVAE,
(SEQ ID NO: 12)
DNVKNKILQIKEEAAKKGVNFKAFTG
TATG GGGNKTFNNLLKLTILVNGGGT
ILVNLLISCGLTGA
(SEQ ID NO: 13)
KDVKNKILQIKKDAEDKGVNFAAFTS
SETG GGGMKKNDQIVAAIALRGVA,
(SEQ ID NO: 14)
KDVKNKILQIKKDAEDKGVNFAAFTS
SETG GGGPFILEAKVRATTVAE,
and
(SEQ ID NO: 15)
KDVKNKILQIKKDAEDKGVNFAAFTS
SETG GGNKTFNNLLKLTILVNGGGTI
LVNLLISCGLTGA,
or an active variant thereof wherein one or more of the amino acids in the linked peptide is substituted with an amino acid replacement, whereby both peptides in the linked peptide specifically bind to an antibody against a pathogenic Borrelia.
2. A diagnostic reagent comprising a composition of claim 1 and a detection system for detecting the peptide, and/or a substrate for immobilizing the peptide.
3. A kit for diagnosing Lyme borreliosis, comprising a composition of claim 1 and a detection system for detecting the peptide bound to an antibody to a pathogenic Borrelia protein and/or a substrate for immobilizing the peptide.
4. A method for diagnosing Lyme disease in a subject, comprising contacting a sample from a subject suspected of having antibodies against a causative agent of Lyme disease with a composition of claim 1 , conditions effective for the formation of a peptide-antibody complex, and detecting the presence of the peptide-antibody complex.
5. The method of claim 4 , wherein the peptide-antibody complex is detected by adding a binding partner which is labeled, or which can be labeled with a signal generating reagent.
6. The method of claim 5 , wherein the binding partner is an antibody attached to an enzyme, and a signal is generated when the enzyme reacts with a suitable substrate.
7. The method of claim 4 wherein the detecting is performed with an ELISA assay.
8. The method of claim 4 , wherein the detecting is performed by immobilizing the peptides onto beads with a unique spectral signature.
9. The method of claim 4 , wherein the subject is a cat or a dog.
10. The method of claim 4 , wherein the subject is a human.
11. The composition of claim 1 , wherein the linked peptide is selected from the group consisting of SEQ ID NOs 6-15.